Protective Effect of Xiangsha Yuyang Decoction on Oxidative Injury in Rats with Gastric Ulcer Based on p62/Keap1/Nrf2 Signal Pathway

Hao ZHANG; Meng-hui CHU; Shi-qi WANG; Zheng-sheng LI; Xue-li SONG; Xiao-ru SHI; Yu-ling WANG

doi:10.13422/j.cnki.syfjx.20210438

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Protective Effect of Xiangsha Yuyang Decoction on Oxidative Injury in Rats with Gastric Ulcer Based on p62/Keap1/Nrf2 Signal Pathway

更新时间：2021-12-03

- Protective Effect of Xiangsha Yuyang Decoction on Oxidative Injury in Rats with Gastric Ulcer Based on p62/Keap1/Nrf2 Signal Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 27, Issue 4, Pages: 56-63(2021)
- 作者机构：
  
  1.河南中医药大学，郑州 450046
  2.河南中医药大学李振华学术思想研究所，郑州 450008
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20210438
  CLC： R2-0;R289;R318.14
- Received：28 November 2020，
  
  Published Online：06 January 2021，
  
  Published：20 February 2021
- 稿件说明：
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张昊,褚梦慧,王诗琦等.基于p62/Keap1/Nrf2信号通路探讨香砂愈疡汤对胃溃疡大鼠氧化损伤的保护作用[J].中国实验方剂学杂志,2021,27(04):56-63.

ZHANG Hao,CHU Meng-hui,WANG Shi-qi,et al.Protective Effect of Xiangsha Yuyang Decoction on Oxidative Injury in Rats with Gastric Ulcer Based on p62/Keap1/Nrf2 Signal Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(04):56-63.
张昊,褚梦慧,王诗琦等.基于p62/Keap1/Nrf2信号通路探讨香砂愈疡汤对胃溃疡大鼠氧化损伤的保护作用[J].中国实验方剂学杂志,2021,27(04):56-63. DOI： 10.13422/j.cnki.syfjx.20210438.

ZHANG Hao,CHU Meng-hui,WANG Shi-qi,et al.Protective Effect of Xiangsha Yuyang Decoction on Oxidative Injury in Rats with Gastric Ulcer Based on p62/Keap1/Nrf2 Signal Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(04):56-63. DOI： 10.13422/j.cnki.syfjx.20210438.

摘要

目的

探讨香砂愈疡汤对乙酸诱导胃溃疡模型大鼠的保护作用并探讨其作用机制，为临床用药提供实验依据。

方法

将60只SPF级Wistar大鼠随机分成6组：空白组，模型组，香砂愈疡汤高、中、低剂量组，奥美拉唑组。通过乙酸诱导制作胃溃疡大鼠模型，香砂愈疡汤高、中、低剂量组分别灌胃28，14，7 g·kg

-1

香砂愈疡汤溶液，奥美拉唑组将奥美拉唑以4.17 g·kg

-1

溶于生理盐水后灌服，空白组和模型组以等体积生理盐水灌胃，1次/d。连续治疗14 d后处死大鼠，采血并取胃组织，测量并计算胃溃疡面积、胃溃疡抑制率，制作胃黏膜组织病理学切片并观察胃黏膜损伤状态；采用酶联免疫吸附测定（ELISA）检测血清中胃黏膜修复因子、胃组织相关蛋白水平、氧化应激因子、炎症因子等指标的变化；采用蛋白免疫印迹法（Western blot）检测胃黏膜组织中p62，Kelch样环氧氯丙烷相关蛋白1（Keap1），核转录因子E2相关因子2（Nrf2），血红素氧合酶-1（HO-1）信号通路相关蛋白的表达水平。

结果

与正常组比较，模型组胃黏膜出现较为明显的病理变化，白细胞大量浸润；模型组溃疡面积显著增加（

0.01），黏蛋白5AC（MUC5AC），表皮生长因子（EGF），超氧化物歧化酶（SOD）和前列腺素E

（PGE

）含量显著减少，胃泌素（GAS），8-羟基脱氧鸟苷酸（8-OHdG），丙二醛（MDA），肿瘤坏死因子-

（TNF-

）和环氧化酶-2（COX-2）含量显著增多（

0.01），Nrf2，HO-1蛋白表达水平显著降低（

0.01），Keap1含量明显增高（

0.05），p62蛋白表达降低

；

与模型组比较，香砂愈疡汤高剂量组和奥美拉唑组细胞层次结构更加清晰，排列规则，形状规整；中、低剂量组对胃黏膜也均有一定的修复作用；香砂愈疡汤高、中剂量组可显著减少乙酸诱导胃溃疡大鼠模型的胃溃疡面积（

0.01），提高溃疡抑制率，明显促进胃黏膜组织中MUC5AC和EGF的表达，降低GAS水平（

0.05，

0.01），显著降低8-OHdG和MDA水平，提高SOD活性（

0.01），显著降低TNF-

和COX-2表达水平，增加PGE

的含量，升高胃黏膜组织Nrf2，HO-1蛋白表达（

0.01），香砂愈疡汤高剂量组可明显降低Keap1的蛋白表达（

0.05），升高p62蛋白表达量。

结论

香砂愈疡汤治疗乙酸诱导胃溃疡模型大鼠效果显著，可有效减少溃疡面积，增加溃疡抑制率，保护溃疡组织。其作用机制可能与激活p62/Keap1/Nrf2信号通路，调控相关基因表达，从而改善炎症反应、调控氧化应激反应有关。

Abstract

Objective

To confirm the protective effect of Xiangsha Yuyang decoction on acetic acid-induced gastric ulcer model rats and explore its mechanism， so as to provide experimental basis for clinical drug use.

Method

The 60 SPF Wistar rats were randomly divided into 6 groups： group， model group， high， middle and low dose groups of Xiangsha Yuyang decoction and omeprazole control group. The rat model of gastric ulcer was induced by acetic acid. The rats in the high， middle and low dose groups of Xiangsha Yuyang decoction were intragastrically administered at the dose of 28，14，7 g·kg

-1

， and with omeprazole at the dose of 4.17 g·kg

-1

in normal saline， respectively. The rats in the blank group and model group were intragastrically infused with the same volume of normal saline once a day. After 14 days of continuous treatment， the rats were killed， the blood was collected， the area and inhibition rate of gastric ulcer were measured and calculated， the histopathological sections of gastric mucosa were made and the state of gastric mucosal injury was observed， and the changes of gastric mucosal repair factor， gastric tissue related protein， oxidative stress factor and inflammatory factor in serum were detected by enzyme-linked immunosorbent assay（ELISA）. Detected the expression of p62 Kelch-like epichlorohydrin-related protein 1 （Keap1）， nuclear transcription factor E2 related factor 2 （Nrf2） and heme oxygenase-1 （HO-1） signal pathway-related proteins in gastric mucosa by Western blot.

Result

Compared with control group， the gastric mucosa of the model group showed obvious pathological changes and a large number of leukocytes infiltrated. In model group， the ulcer area was significantly increased（

0.01）， the contents of mucin mucoprotein 5AC （MUC5AC）， epidermal growth factor （EGF）， superoxide dismutase （SOD） and increased prostaglandin E

（PGE

） were significantly decreased（

0.01）， the gastrin （GAS）， 8-hydroxydeoxyguanosine （8-OHdG）， malondialdehyde （MDA）， tumor necrosis factor-

（TNF-

）， cyclooxygenase-2 （COX-2） were significantly increased. The expression of HO-1 and Nrf2 protein decreased significantly（

0.01）， the content of Keap1 increased significantly （

0.05）， and the expression of p62 protein decreased. Compared with model group， the hierarchical structure of cells in Xiangsha Yuyang decoction high dose group and omeprazole group were clearer and regular， middle and low dose groups could also repair gastric mucosa to a certain extent. The high and middle dose groups of Xiangsha Yuyang decoction could significantly reduce the gastric ulcer area of acetic acid-induced gastric ulcer rat model （

0.01） and increase the ulcer inhibition rate. It can effectively promote the expression of MUC5AC and EGF in gastric mucosa， decrease the level of GAS（

0.05，

0.01）， decrease the level of 8-OHdG and MDA， increase the activity of SOD（

0.01）， decrease the expression level of TNF-

and COX-2， increase the content of PGE

， and significantly increase the amount of Nrf2 and HO-1 protein in gastric mucosa（

0.01）. The high dose group of Xiangsha Yuyang decoction could decrease the protein expression of Keap1（

0.05） and increase the expression of p62 protein.

Conclusion

Xiangsha Yuyang decoction is effective in the treatment of acetic acid-induced gastric ulcer model rats， which can effectively reduce the ulcer area， increase the ulcer inhibition rate and protect the ulcer tissue. Its mechanism may be related to activating p62/Keap1/Nrf2 signal pathway and regulating the expression of related genes so as to improve inflammatory response and regulate oxidative stress response.

关键词

Keywords

references

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